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A dual vaccine against avian metapneumovirus and h9 subtype avian influenza virus

A technology for avian metapneumovirus and avian influenza virus, applied in the direction of viruses, vaccines, antiviral agents, etc., can solve the problem of ineffective prevention of avian metapneumovirus and H9 subtype avian influenza, low antibody level and uniformity of antibodies, affecting vaccines Prevention and control effects and other issues, to achieve the effect of improving uniformity, increasing antibody levels, and good safety

Active Publication Date: 2020-11-27
YEBIO BIOENG OF QINGDAO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The existing avian influenza (H9 subtype) vaccine has been widely used clinically, but there are problems with low antibody levels and antibody uniformity, which affect the prevention and control effect of the vaccine
However, there is currently no effective dual vaccine against avian metapneumovirus and H9 subtype avian influenza in the market

Method used

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  • A dual vaccine against avian metapneumovirus and h9 subtype avian influenza virus
  • A dual vaccine against avian metapneumovirus and h9 subtype avian influenza virus
  • A dual vaccine against avian metapneumovirus and h9 subtype avian influenza virus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1: Amplification and sequence analysis of F protein

[0022] In 2010, symptoms of swollen head syndrome appeared in several breeding chicken farms in Shandong Province, and the affected chickens had been injected with the existing avian pneumovirus vaccine before, and it was speculated that the infected virus had mutated; Screening for lung viruses. Finally, an avian pneumovirus SHS / A3 was screened out.

[0023] In order to verify the antigenicity of the screened virus, virus strains from 5 different sources, including the screened strain SHS / A3, were used as antigens to prepare vaccines. After immunizing SPF chickens, the SHS / A3 strain virus solution was used to challenge the virus. The results It shows that compared with other poultry pneumovirus vaccines; the vaccine prepared by itself has a better immune effect (p<0.05); therefore, it is determined that genetic variation has occurred.

[0024] 1. Amplification of Type B Avian Metapneumovirus SHS / A3 Strain ...

Embodiment 2

[0035] Embodiment 2: Recombinant expression of F protein

[0036] 1. The preparation method of recombinant type B avian metapneumovirus F protein

[0037] The method comprises the following steps: a. constructing an expression vector; b. constructing an expression strain; c. inducing, extracting and purifying the recombinant F protein.

[0038] a. Construct expression vector:

[0039] The positive clone plasmid pMD18-T-F and the expression vector pPICZα vector were double-digested with KpnI and NotI respectively, electrophoresed on a 1.2% agarose gel, and then recovered with a DNA gel recovery kit to obtain about 1.6kb and 3.3kb fragments, respectively. The pPICZα-F expression vector was constructed by directional ligation at 16°C ( Figure 4 It is the enzyme digestion identification diagram of the high-efficiency expression vector of the embodiment of the present invention); after sequencing and verifying that the sequence and reading frame are correct, the plasmid is linea...

Embodiment 3

[0044] Embodiment 3: Preparation of antigen for seedling production

[0045] 1. Preparation of avian influenza virus liquid for making seedlings Inoculate 10-11-day-old SPF chicken embryos through the allantoic cavity with the preservation number CCTCC NO:V201517 H9 subtype avian influenza virus QDY strain, 0.2ml per embryo, 36°C Incubate at ~37°C, take embryos twice a day, take dead chicken embryos after 24 hours, and harvest them at 96 hours, regardless of whether they are dead or not, cool at 4-8°C for 12-24 hours, harvest chicken embryo liquid, mix in sterile Store in a container at 2-8°C. The virus content of the harvested virus liquid is determined, and the virus content per 0.1ml is not less than 10 8.0 EID 50 ;

[0046]2. Inactivation Put the qualified venom into a sterilization container, add 10% formalin solution to a final concentration of 0.2%, and inactivate at 37°C for 18 hours;

[0047] 3. Preparation of F protein of avian metapneumovirus type B for seedling...

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Abstract

The invention provides a bivalent vaccine of an acian metapneumovirus and an H9subtype avian influenza virus. The bivalent vaccine comprises an antigen and an vaccine adjuvant, wherein the antigen is an H9 subtype avian influenza virus strain and acian metapneumovirus F protein; thepreservation numberof the H9 subtype avian influenza virus strain is CCTCC NO.V201517; and the amino acid sequence of the acian metapneumovirus F protein is SEQ ID NO:1. The H9 subtype avian influenza virus strain QDY and the acian metapneumovirus F protein antigen solution are mixed at the ratio, and then an oil adjuvant is added to the mixture to be mixed and emulsified into the vaccine. According to the prepared vaccine, the immunized antibody level can be improved, the uniformity of the immunized antibody is improved, the immune effect of the vaccine is ensured, and the vaccine has the advantages of high efficiency and good safety.

Description

technical field [0001] The invention belongs to the technical field of veterinary biological products, and in particular relates to a dual vaccine of avian metapneumovirus and H9 subtype avian influenza virus. Background technique [0002] Avian metapneumovirus belongs to the paramyxovirus pneumovirus genus, and its genome is a single-stranded negative-sense RNA without segments, which can cause respiratory diseases in turkeys, called turkey rhinotracheitis; after infecting chickens, it can cause viral air sacculitis, Rhinotracheitis and swollen head syndrome, decreased egg production. In the late 1970s, the disease caused by avian metapneumovirus was first reported in South Africa, and subsequently occurred in North America, South America, the Middle East, and the Far East. The virus was first isolated in the United States in 1989. While isolating the virus, it is often possible to isolate Bordetella, Pasteurella, Pseudomonas, Rhinotracheal Avian bacillus, and Alcaligenes...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/155A61P31/14A61P31/16A61K39/145
CPCA61K39/12A61K2039/521A61K2039/70C12N2760/16134C12N2760/18334
Inventor 刘新文王秀丽邹敏郭伟伟宫晓王龙邹桂荣
Owner YEBIO BIOENG OF QINGDAO
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